A presentation on Exporting the Prospect of RSV Vaccine for Infants with Cardiovascular Disease Through Gene Modification by Jesse L. Ramey, an Avalon University School of Medicine Basic Science student.

1. Exploring the Prospect of RSV
Vaccine for Infants With
Cardiovascular Disease Through
Gene Modification
PRESENTED BY JESSE L. RAMEY

2. RSV causes…
2,000,000 hospitalizations of
infants in the US [1]
200,000 deaths/yr. of infants
in developing countries [2]
Infants with cardiovascular
disease are especially
susceptible
RSV forms a syncytium of infected in vitro cells.
Background:

3. Methods: Study effect of mutating F & G proteins by
removing gycans on RSV infection [4], [5]

4. Methods (continued):
1.) Sever the gene of interest from the RSV genome with restriction enzymes.
2.) Alter the nucleotide sequence for the O-linked glycan on the F protein by changing the ACC sequence
for threonine to ATG for methionine to lose the O-linked glycan from the F protein.
3.) Replicate with recombinant DNA

5. Methods (continued):
Isolate, sequence and select the best DNA to be used for
transfection of HEK-280 cells.
Sample #4 produced the best DNA.

6. Final Steps of method:
1.) Transfect HEK-280 cells with DNA.
2.) Culture the HEK-280 cells.
3.) Observe cells for syncytium.
4.) Lyse cells and analyze protein with a Western Blot.
5.) Compare to WT protein for verification of glycan of interest.

7. Results:
Syncytium with WT F protein but no syncytium formed with mutated protein.
O-linked glycan of interest contributes to infection.
The Western Blot was inconclusive

8. Conclusion:
A beta version of the vaccine is functional in the laboratory with the cotton
mouse, and research teams seek to strengthen this vaccine with additional
genetic modifications made possible by testing different aspects of RSV
proteins.
This research is still in progress.
It is contributory to the process of using genetically modified RSV as a
vaccine.

9. References:
 [1] CDC.gov, Respiratory Syncytial Virus Infection, Available from
https://www.cdc.gov/rsv/research/us-surveillance.html [Accessed July 8, 2017]
 [2] path.org, RSV Disease, Available from http://www.path.org/vaccineresources/rsv.php
[Accessed July 8, 2017]
 [3] Wn Jung, J, Respiratory syncytial virus infection in children with congenital heart
disease: global data and interim results of Korean RSV-CHD survey, Korean Journal of
Pediatrics. 2011; 54(5): 192-196.
 [4] McLellan JS1, Ray WC, Peeples ME., Structure and function of respiratory syncytial
virus surface glycoproteins., Current Topics Microbiology Immunology. 2013;372:83-104.
doi: 10.1007/978-3-642-38919-1_4.]
 [5] McDonald TP, Jeffree CE, Li P, Rixon HW, Brown G, Aitken JD, MacLellan K, Sugrue RJ.,
Evidence that maturation of the N-linked glycans of the respiratory syncytial virus (RSV)
glycoproteins is required for virus-mediated cell fusion: The effect of alpha-mannosidase
inhibitors on RSV infectivity, Virology. 2006;350(2):289-301.

10. Acknowledgements:
 Mr Abraham Ratna Joseph N , Avalon University
 Dr Yogesh Acharya, Avalon University
 Dr Sireesha Bala Arja, Avalon University
 Dean of Basic Sciences, Avalon University
 Dr Sateesh B Arja, Executive Dean, Avalon University
 Dr Samir Fatteh President, Avalon University
 Dr Shokat Fatteh Chancellor., Avalon University
 Dr. Mark Peeples, Nationwide Children’s Hospital

11. THANK YOU